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[要旨]ペンギン類は、同種内で雌雄個体間に外見の差が少ないため、外見からは個体の雌雄を正確に判別することが困難である。その一方で、日本国内で飼育され、動物園・水族館で展示されているペンギン類を維持し、展示を継続するためには、国内での効率の良いペンギン類の繁殖が必要であり、その第一段階として、ペンギン類の各個体の性別を知ることは必須である。本稿では、当研究室で実施しているPCR 法によるペンギン類の雌雄判別について紹介する。[英文抄録] Since there is little difference in appearance between males and females, it is difficult to accurately distinguish the sex of each individual penguins from the appearance. On the other hand, in order to maintain and continue the exhibition of penguins bred at zoos and/or aquariums in Japan, efficient breeding of penguins is necessary. As a step of efficient breeding, it is essential to know the sex of each individual penguins. In this article, I will introduce the sexing of penguins by the PCR method carried out in my laboratory., Research Notes

In mammals, sexual fate is determined by the chromosomes of the male and female gametes during fertilization. Males (XY) or females (XX) are produced when a sperm containing a Y or X-chromosome respectively fertilizes an X-chromosome-containing unfertilized egg. However, sexing of preimplantation stage embryos cannot be conducted visually. To address this, transgenic male mouse models with the ubiquitously expressed green fluorescent protein (GFP) transgene on X- (X-GFP) or Y-chromosomes (Y-GFP) have been established. However, when crossed with wild-type females, sexing of the preimplantation stage embryos by observing the GFP signal is problematic in some cases due to X-inactivation, loss of Y-chromosome (LOY), or loss of transgene fluorescence. In this study, a mouse model with the ubiquitously expressed red fluorescent protein (RFP) transgene on the Y-chromosome was generated since RFP is easily distinguishable from GFP signals. Unfortunately, the ubiquitously expressed tdTomato RFP transgene on the Y-chromosome (Y-RFP) mouse showed the lethal phenotype after birth. No lethal phenotypes were observed when the mitochondrial locating signal N-terminal of tdTomato (mtRFP) was included in the transgene construct. Almost half of the collected fertilized eggs from Y-mtRFP male mice crossed with wild-type females had an RFP signal at the preimplantation stage (E1.5). Therefore, XY eggs were recognized as RFP-positive embryos at the preimplantation stage. Furthermore, 100% sexing was observed at the preimplantation stage using the X-linked GFP/Y-linked RFP male mouse. The established Y-mtRFP mouse models may be used to study sex chromosome related research.

The present trial explained the effect of alternative production systems on growth, morphometric and carcass traits of four different chicken genotypes. The second generation of two genotypes RNN (Rhode Island Red × Naked Neck) and BNN (Black Australorp × Naked Neck) obtained by two self-crosses (RNN × RNN = RR and BNN × BNN = BB) and two reciprocal crosses (RNN × BNN = RB and BNN × RNN = BR) were evaluated in three alternative production systems (conventional cages, enriched cages, and aviary). At the 6th week of age after sexing, a total of 600 birds, comprising 150 from each crossbred with a total of 300 pullets and 300 cockerels were divided into conventional cages, enriched cages, and aviary systems having 200 birds in each.Birds were organized into 3×4 factorial arrangements under Completely Randomized Design (3 production systems × 4 genotypes × 2 sexes × 25 birds = 600 birds). Regarding genotypes, RB and BR males showed higher (p < 0.01) carcass yield, drumstick weight, breast weight, and thigh weight than BB and RR genotypes. Females of BR genotype showed higher (p < 0.01) breast weight, thigh weight and drumstick weight. As far as production systems are concerned, higher (p < 0.01) liver weight, heart weight, breast weight, intestinal weight, drumstick weight, and thigh weight were observed in the males reared in enriched cages compared with conventional cages and aviary system. Females reared in enriched cages showed higher (p < 0.01) heart weight, breast weight, intestinal weight, drumstick weight, and thigh weight when compared with those reared in conventional cages and aviary. It is concluded that chickens (both sexes) of BR and RB genotypes had better morphological measurements and carcass traits than those of RR and BB genotype chickens. Among alternative production systems, chickens reared in enriched cages had better traits than those of reared in conventional cages and aviary during the growing phase. Resumo O presente estudo explicou o efeito de sistemas alternativos de produção sobre o crescimento, características morfométricas e carcaças de quatro genótipos de frango diferentes. A segunda geração de dois genótipos RNN (Rhode Island Red × Naked Neck) e BNN (Black Australorp × Naked Neck) obtida por duas autocruzes (RNN × RNN = RR e B ANN × BNN = BB) e duas cruzes recíprocas (RNN × BNN = RB e BNN × RNN = BR) foi avaliada em três sistemas de produção alternativos (gaiolas convencionais, gaiolas enriquecidas e aviário). Na 6ª semana de idade após o sexo, um total de 600 aves, compostas por 150 de cada raça cruzada com um total de 300 pullets e 300 galos, foi dividido em gaiolas convencionais, gaiolas enriquecidas e sistemas aviários com 200 aves em cada. As aves foram organizadas em 3×4 arranjos fatoriais sob projeto completamente randomizado (3 sistemas de produção × 4 genótipos × 2 sexos × 25 aves = 600 aves). Em relação aos genótipos, os machos RB e BR apresentaram maior rendimento de carcaça (p < 0,01), peso da baqueta, peso mamário e peso da coxa do que os genótipos BB e RR. As fêmeas do genótipo BR apresentaram maior (p < 0,01) peso mamário, peso da coxa e peso da baqueta. No que diz respeito aos sistemas de produção, maior (p < 0,01) peso hepático, peso cardíaco, peso mamário, peso intestinal, peso da baqueta e peso da coxa foram observados nos machos criados em gaiolas enriquecidas em comparação com gaiolas convencionais e sistema aviário. As fêmeas criadas em gaiolas enriquecidas apresentaram maior (p < 0,01) peso cardíaco, peso mamário, peso intestinal, peso da baqueta e peso da coxa quando comparadas com as criadas em gaiolas convencionais e aviárias. Conclui-se que as galinhas (ambos os sexos) dos genótipos BR e RB apresentaram melhores medidas morfológicas e traços de carcaça do que os de frangos genótipos RR e BB. Entre os sistemas de produção alternativos, as galinhas criadas em gaiolas enriquecidas tinham características melhores do que as criadas em gaiolas convencionais e aviárias durante a fase de cultivo.

Sex-specific ecology has management implications, but rapid sex-chromosome turnover in fishes hinders sex-marker development for monomorphic species. We used annotated genomes and reduced-representation sequencing data for two Australian percichthyids, Macquarie perch Macquaria australasica and golden perch M. ambigua, and whole genome resequencing for 50 Macquarie perch of each sex, to identify sex-linked loci and develop an affordable sexing assay. In-silico pool-seq tests of 1,492,004 Macquarie perch SNPs revealed that a 275-Kb scaffold was enriched for gametologous loci. Within this scaffold, 22 loci were sex-linked in a predominantly XY system, with females being homozygous for the X-linked allele at all 22, and males having the Y-linked allele at >7. Seven XY-gametologous loci (all males, but no females, are heterozygous or homozygous for the male-specific allele) were within a 146-bp region. A PCR-RFLP sexing assay targeting one Y-linked SNP, tested in 66 known-sex Macquarie perch and two of each sex of three confamilial species, plus amplicon sequencing of 400 bp encompassing the 146-bp region, revealed that the few sex-linked positions differ between species and between Macquarie perch populations. This indicates sex-chromosome lability in Percichthyidae, supported by non-homologous scaffolds containing sex-linked loci for Macquarie- and golden perches. The present resources facilitate genomic research in Percichthyidae, including formulation of hypotheses about candidate genes of interest such as transcription factor SOX1b that occurs in the 275-Kb scaffold ~38 Kb downstream of the 146-bp region containing seven XY-gametologous loci. Sex-linked markers will be useful for determining genetic sex in some populations and studying sex chromosome turnover.

Incorporating sex ratios of nestlings into population viability studies increases knowledge of overall health of endangered populations. Currently, a reliable non-invasive method to identify the sex of golden eagle nestlings is not available; however, claims are commonly made based on morphology. Ten biometric measurements from 43 Scottish golden eagles aged 2–7.5 weeks were assessed to see if sex could actually be determined using this non-invasive methodology. Sex was confirmed via molecular analysis of blood samples. Discrete and principal component analyses of the different biometrics could not correctly determine individual nestling sex. Therefore, despite being more invasive, molecular sexing remains the recommended tool of choice for accurate sex identification of Scottish golden eagle nestlings younger than 7.5 weeks of age. This has important implications for golden eagle field studies where empirical morphological measurements are frequently and typically taken, but we have shown are not reliable in determining the sex of such young nestlings.

A lo largo de cinco secciones, se recapitulan las principales biotecnologías reproductivas en el bovino, se analiza su desarrollo histórico, estado actual, y se incluyen resultados generados en México. En la década de 1970, se inició la sincronización estral e inducción de la ovulación donde, basados en el conocimiento de la fisiología reproductiva bovina, se empezó a controlar el ciclo reproductivo con recursos disponibles en aquel entonces. Con los años, la terapia hormonal evolucionó conforme se descubrieron nuevos compuestos, refinando métodos para estandarizar el efecto y generar nuevos métodos de liberación de las hormonas. La biotecnología más usada en el mundo, la inseminación artificial, debe su expansión a los avances en el procesamiento del semen, donde destaca el desarrollo de diluyentes, la criopreservación, el sexado del semen y el análisis espermático asistido por computadora. La era embrionaria inició con el desarrollo de la multiovulación y los métodos para colectar, evaluar, transferir y criopreservar los embriones. La segunda era embrionaria llegó con la producción de embriones completamente in vitro, partiendo de óvulos inmaduros y semen congelado, en lo que se denominó la producción in vitro de embriones. En ésta, se han invertido grandes recursos de investigación, y materiales, para hacerla un pilar del mejoramiento genético y la productividad, en combinación con otras dos herramientas, el semen sexado y las evaluaciones genómicas. Se vislumbra una época de oro de la producción in vitro de embriones con la posibilidad de modificar el genoma de embriones con precisión, gracias a la tecnología de edición de genes.

Background and Aim: Lecithin based diluent such as AndroMed, is a semen diluent made without animal components to prevent the risk of disease transmission, while glutathione (GSH) is an intracellular non-enzymatic antioxidant that prevents cell damage due to reactive oxygen species. Meanwhile, the specific impact of AndroMed and GSH combinations on spermatozoa abnormalities has not been fully studied. Therefore, this study aims to determine the effect of using cauda epididymal plasma-2 (CEP-2) and AndroMed diluents with or without the addition of GSH on the abnormalities of sexing semen of Ongole crossbred bulls in cold storage. Materials and Methods: This study used a factorial completely randomized design 2×2, the first factor was types of diluent and the second was with or without the addition of GSH. Observation of spermatozoa abnormalities was carried out at a storage time of 0-5 days using 297 ejaculations of liquid semen, with 100 spermatozoa observed per smear of each ejaculate. Furthermore, the data were analyzed using a two-way analysis of variance and the significant threshold (p-value) for statistical analysis was set at

Documenting the sex of individuals encountered during wildlife research and monitoring activities is important for understanding and tracking changes in populations. However, sexing salamanders can be particularly difficult because secondary sex characters are often subtle or only visible during the breeding season, and guidance on species-specific sex determination is lacking from most field guides. The purpose of this guide is to provide a reference to assist biologists in the Central Appalachian region with identifying sex of live adult salamanders. In the main text we provide summary tables and figures to serve as concise references in the field. We provide individual species accounts that contain concise yet comprehensive information for each species based on the published literature, as well as many images depicting sexually dimorphic characters. Our focal region encompasses partial or entire distributions for 56 species of salamanders in 5 families (Ambystomidae, Cryptobranchidae, Plethodontidae, Proteidae, and Salamandridae). We identified seven morphological characters that are strongly sexually dimorphic and useful for sexing live, nonanesthetized, adult salamanders in the field, with males of individual species exhibiting one to five of the characters. We identified more than 20 additional characters that are weakly sexually dimorphic, difficult to distinguish in the field, or species-specific. Our guide serves as a synthesis of sexually dimorphic characters for salamanders in Central Appalachia, and we anticipate it will have broad value for researchers, monitoring programs, and salamander enthusiasts in eastern and central North America.

Hormonal sex - reversal is widely recognized as having significant advantage over both manual sexing and hybridization. The present study carried out in the aquaculture center of Kankan aimed at producing monosex male fry of Oreochromis niloticus in order to increase the yield of this species. During this study, 95 Oreochromis niloticus broodstock including 21 males and 74 females were paired (spawned) in a spawning tank of 135 m2 surface for 15 days. The respective mean weights of the spawners were 448.6 ±157.7 g for males and 406.7±115.9 g for females. Hormonal treatment was performed on 900 larvae with an average initial weight of 0.028 g from the 10th post fertilization to the 38th day in a tank of one (1) m3. The larvae were fed with 1 kg of imported "Raanan Premium Fish" powdered feed titrated with 40% protein, mixed in a hormonal solution containing 17-α-methyltestosterone and 90% ethanol with respective doses of 100 mg and 500 ml. At the end of hormonal treatment, 578 fry with a final average weight of 4.5 g were conducted in a pre-pregnancy tank with a surface area of 50 m2 for 30 days and yielded at the end of pre-pregnancy 501 fry with a final average weight of 13 g, a daily growth rate of 0.22 g, a specific growth rate of 1.7%, a survival rate of 64% and a mortality rate of 36%. After sex identification (sexing), the rates of hormonal treatment were 83.33% and 16.66% for males and females, respectively. In conclusion, the popularization of the results of this study could be considered as an alternative for a production of monosex male population in Oreochromis niloticus that will feed the networks of local producers to improve the production of the species throughout the country. Keywords: Oreochromis niloticus, hormone, sex, larvae and tank.

Background Sexual differentiation can occur after exposure to aromatase into the left gonad at 6.5 days of incubation. Aromatase inhibitors work by inhibiting the action of the aromatase, which converts androgens into estrogens by a process called aromatization. Objectives The aim of this study was to investigate the effect of in ovo exposure to the aromatase inhibitor from tomato and garlic extract on sexual differentiation and embryonic development in chicken embryos. Methods Three hundred eggs divided into five groups: Control 1 (CO; no injection); control 2 distilled water, DW; 0.1 ml/egg); garlic extract (GAR; 0.1 mg/egg); tomato extract (TOM; 0.1 mg/egg); and garlic and tomato extract mixed (ATM, 0.1 ml/egg). The solution was prepared and injected into the albumin from the thin end of the eggs on day five by using a 1 ml syringe with a 23‐gauge needle. The embryonic test (embryo/egg weight) conducted at 7, 14 and 17 days of incubation. After hatching, feather sexing conducted to determine the initial male. Chicks sex was later confirmed on day 42 by an optical microscope lens. Results The results revealed that there was a significant increase (p, The aim of this study was to investigate the effect of in ovo exposure to the aromatase inhibitor of tomato and garlic extract on sexual differentiation and embryonic development of chicken embryos. Sexual differentiation can occur after exposure to aromatase into the left gonad at 6.5 days of incubation. Aromatase inhibitors work by inhibiting the action of the aromatase, which converts androgens into estrogens by a process called aromatization.

The Eurasian tree sparrow (Passer montanus) is a monomorphic passerine, for which it is impossible to differentiate between males and females based on external characteristics. Being a species frequently captured for ringing, having a reliable method to determine sex from conventional biometric measurements would facilitate its study and be very useful for the correct management and conservation of this declining species. In the present study, we used biometric measurements recorded in 66 individuals captured with mist nets in communal roosts in northern Spain during the winter and sexed them using molecular techniques. We conducted a discriminant function analysis (DFA) to derive equations that allowed us to determine the sex of the specimens from some of the measurements recorded in the field. Significant differences were found between males and females in wing length, third primary length, badge width and height and body weight. The DFA provided two functions that correctly classified the sex of 94.7% of the individuals using wing length and badge width, and 98.2% if weight was added to the analysis. Our results allow sexing from measurements that can be easily recorded in the field with the tools commonly used in banding sessions and without the need for additional training. Considerations of Bergmann’s and Allen’s rules on body size and the use of DFA in different populations are discussed.

International audience; Chub mackerel Scomber japonicus and blue mackerel S. australasicus have become important aquaculture target species in Japan. For an efficient and stable aquaculture production of mackerels, it is desirable to work with a female-biased broodstock and thus to be able to manage precisely their sex ratio. As these two mackerel species do not have any external sexual dimorphism, direct biopsy of the gonads or measurement of sex hormone plasma levels is used in research to determine their phenotypic sex. However, these methods are associated with handling stress that results in gonadal atresia. Besides, it is difficult to identify sex using these methods in immature individuals. Therefore, genetic sexing would be desirable in these species to overcome these problems. In this study, we developed molecular sex identification tools for both chub and blue mackerels using sex-specific single-nucleotide variations (SNVs) identified by pool-sequencing (pool-seq) analyses. Genomic DNA was isolated from 30 males and females, in each species, and sequenced as pooled samples. Results of these pool-seq analyses identified a clear XX/XY male heterogametic sex-determination system in blue mackerel and a ZZ/ZW female heterogametic system in chub mackerel. Using this pool-seq information, we designed primer sets for genetic sex identification in both species. Results demonstrated that these primer sets are robust and accurate in assessing the genotypic sex of both blue and chub mackerels (accuracy > 99.5% in the chub, and > 94.5% in the blue mackerel) in various geographical samples of different ages. This method can now be applied for the efficient management of sex ratios in both mackerel aquaculture populations and wild-caught animals for fisheries management.

Background Goat milk is very similar to human milk in terms of its abundant nutrients and ease of digestion. To derive greater economic benefit, farmers require more female offspring (does); however, the buck-to-doe offspring sex ratio is approximately 50%. At present, artificial insemination after the separation of X/Y sperm using flow cytometry is the primary means of controlling the sex of livestock offspring. However, flow cytometry has not been successfully utilised for the separation of X/Y sperm aimed at sexing control in dairy goats. Results In this study, a novel, simple goat sperm sexing technology that activates the toll-like receptor 7/8 (TLR7/8), thereby inhibiting X-sperm motility, was investigated. Our results showed that the TLR7/8 coding goat X-chromosome was expressed in approximately 50% of round spermatids in the testis and sperm, as measured from cross-sections of the epididymis and ejaculate, respectively. Importantly, TLR7/8 was located at the tail of the X-sperm. Upon TLR7/8 activation, phosphorylated forms of glycogen synthase kinase α/β (GSK3 α/β) and nuclear factor kappa-B (NF-κB) were detected in the X-sperm, causing reduced mitochondrial activity, ATP levels, and sperm motility. High-motility Y-sperm segregated to the upper layer and the low-motility X-sperm, to the lower layer. Following in vitro fertilisation using the TLR7/8-activated sperm from the lower layer, 80.52 ± 6.75% of the embryos were XX females. The TLR7/8-activated sperm were subsequently used for in vivo embryo production via the superovulatory response; nine embryos were collected from the uterus of two does that conceived. Eight of these were XX embryos, and one was an XY embryo. Conclusions Our study reveals a novel TLR7/8 signalling mechanism that affects X-sperm motility via the GSK3 α/β-hexokinase pathway; this technique could be used to facilitate the efficient production of sexed dairy goat embryos.

Both English writer Jeanette Winterson and Turkish writer Elif Shafak are marked through their involvement with feminism. Considering their concern with mysticism as a reaction against the Orthodox religion which they believe gives support to the patriarchal oppression of women, this study compares Winterson’s Sexing the Cherry and Shafak’s Pinhan. Winterson deals with Kabbala mysticism in Sexing the Cherry whereas Shafak adapts Sufism in Pinhan. Both authors depict, in their novels, fluid character identities who reincarnate in different times and spaces in different bodies. This attitude is considered, even though they adapt different types of mysticism, as the reflection of their concern with spirituality, which they develop against the discriminating patriarchal religion. By privileging spiritual reality over the physical reality, they both annihilate women’s subjection to physical laws of patriarchy.

The use of sexed semen in dairy and beef farms ensures the production of animals of the desired sex, resulting in a reduction of costs and an improvement of environmental sustainability. Several methods have been developed over the years, but most of them were abandoned due to their limited efficacy. Currently, the only commercially available method for the separation of X‐ and Y‐chromosome‐bearing sperm is fluorescence‐activated cell sorting. However, this technique is expensive and has limited usefulness for the industry, considering that it cannot produce doses of sexed semen with the desired number of sperm for artificial insemination. Immunological methods have emerged as an attractive alternative to flow cytometry and proteomic knowledge of X‐ and Y‐sperm could be useful to the development of a new method. In this review, we identify the main applications of sexed semen, describe the existing methods and highlight future research opportunities in the field. We consider that immunological methods, based on sperm cell's surface proteins differentially expressed between X‐ and Y‐sperm, could be an interesting and promising approach to semen sexing., Immunological methods have emerged as an attractive alternative to flow cytometry and proteomic knowledge of X‐ and Y‐sperm could be useful to the development of a new method. In this review we identify the main applications of sexed semen, describe the existing methods and highlight the future research opportunities in the field. We consider that immunological methods, based on sperm cell's surface proteins, deferentially expressed between X‐ and Y‐sperm could be an interesting and promising approach to sex selection.

A research group of Animal Science Faculty of Brawijaya University introduced and implemented Frozen sexed semen Artificial Insemination to 120 beef cattle farmers in Palang Village, Tuban Regency, East Java, from 2017 to 2019. The result of the program reported that the success rate of the technology is quite high, with the birth rate of male claves up to 80%. However, farmers’ adoption rate of the technology was relatively low. This research aims to evaluate factors that affected the adoption rate including the technology, characteristics of respondents, and the extension agents. Primary data were collected through a census of 120 farmers involved in the program and in-depth interviews with stakeholders. Secondary data were collected through related documents including reports of the programs, Statistics, and other related documents. This research utilized Quantitative and qualitative analyses. Data of the study indicated that respondents have neutral perceptions of the Frozen Semen Sexing Artificial Insemination. Although male calves relatively have a higher price, more than 50% of respondents did not expect specific sexing for the calves. In-depth interview data revealed that the success of the Artificial insemination technology is more valuable for the farmers rather than the sexing. Characteristics of respondents and the extension agents played significant roles in the adoption rate of the Frozen Sexed Semen Artificial Insemination technology adoption in Palang Village, Tuban Regency, East Java.

Zophobas morio, es un Coleoptera, Tenebrionidae, de color negro opaco, robusto y de rápido desplazamiento. Son sus larvas y no los adultos, quienes resultan muy útiles en la alimentación de animales como peces, arañas, aves, reptiles, escorpiones, anfibios y mamíferos, por su alto contenido de proteínas. Este es uno de los motivos que justifica su importancia e incentiva su cría con fines comerciales y de experimentación. En varios ensayos con este insecto, es necesario formar parejas de macho y hembra, lo que puede hacerse tanto en la etapa de pupa como en la de adulto. Este trabajo tiene como finalidad mostrar por medio de imágenes, el dimorfismo sexual de Z. morio tanto en la etapa de pupa, como en la adulta, con ejemplares vivos y muertos. Los adultos muertos fueron clarificados en KOH al 10% para facilitar su observación. Las imágenes son fotografías de tipo descriptivas y por el enfoque didáctico de la investigación, se elaboraron dos mapas conceptuales para resumir por pasos el trabajo realizado e incentivar la repetibilidad. Las fotografías de pupas y adultos se realizaron con un microscopio estereoscópico Wild M7A y una cámara digital Panasonic Lumix, DMC-ZS20. Se obtuvo como resultado de las observaciones, una diferencia notable en la forma de las papilas genitales de la pupa hembra, con respecto a las del macho. Igualmente al comparar el borde de la región frontoclipeal del adulto macho con el de la hembra, se evidenció claramente el dimorfismo sexual. La metodología utilizada fue bastante acertada para sexar al insecto en dos fases de su ciclo biológico. Así, además de las claves taxonómicas existentes, este trabajo es un apoyo fotográfico y didáctico de utilidad en la identificación de la especie.

Differential epigenetic (DNA cytosine methylation) and gene expression patterns were investigated in reproductive and vegetative organs from Ilex paraguariensis and I. dumosa, at distinct developmental stages. We aimed at contributing towards elucidating major molecular changes underlying the sexual differentiation processes which, in these dioecious species, are completely unknown. Simultaneously, as a first step towards the development of an early sexing system, we searched for promising molecular markers. This was assessed through Methylation Sensitive Amplified Polymorphism (MSAP) and Amplified Fragment Length Polymorphism on cDNA (cDNA-AFLP) techniques, applying discriminant multivariate analyses, and bioinformatic characterization of differential fragments. A significant positive correlation was found between epigenetic and indirect 'genetic' information for both species, indicating influence of the genetic background on the epigenetic variation. Higher epigenetic than genetic diversities were estimated. Our outcomes showed up to 1.86 times more representation of mCG subepiloci than mCCG in all organs sampled. Along the maturing stages of floral buds, the frequency of mCG evidenced an incremental trend, whereas mCCG and unmethylated conditions showed opposite tendencies. Reproductive and vegetative samples tended to cluster apart based on epigenetic patterns; at gene expression level, organs exhibited clear-cut distinctive patterns, nonetheless profiles of young leaves and floral primordia resemble. Epigenetic and expression data allowed discrimination of I. dumosa´s samples according to the gender of the donor; more elusive patterns were observed for I. paraguariensis. In total, 102 differentially methylated and expressed fragments were characterized bioinformatically. Forty-three were annotated in various functional categories; four candidate markers were validated through qPCR, finding statistical differences among organs but not among sexes. The methylation condition of epilocus C13m33 appears as indicative of gender in both species. Thirty-three organ-specific and 34 gender-specific methylated markers were discriminated and deserve further research, particularly those expressed in leaves. Our study contributes concrete candidate markers with potential for practical application.

Gene expression analysis in preimplantation embryos has been used for answering fundamental questions related to development, prediction of pregnancy outcome, and other topics. Limited amounts of mRNA in preimplantation embryos hinders progress in studying the preimplantation embryo. Here, a method was developed involving direct synthesis and specific-target preamplification (STA) of cDNA for gene expression analysis in single blastocysts. Effective cell lysis and genomic DNA removal steps were incorporated into the method. In addition, conditions for real-time PCR of cDNA generated from these processes were improved. By using this system, reliable embryo sexing results based on expression of sex-chromosome linked genes was demonstrated. Calibration curve analysis of PCR results using the Fluidigm Biomark microfluidic platform (Fluidigm, South San Francisco, CA) was performed to evaluate 96 STA cDNA from single blastocysts. In total, 93.75% of the genes were validated. Robust amplification was detected even when STA cDNA from a single blastocyst was diluted 1,024-fold. Further analysis showed that within-assay variation increased when cycle threshold values exceeded 18. Overall, STA quantitative real-time PCR analysis was shown to be useful for analysis of gene expression of multiple specific targets in single blastocysts.

Pangasius (P. hypophthalmus) is one of the freshwater commodities of an important economic value. The pH on fertilization media can influence fertilization and affect sex determination (Sandra and Norma, 2009). Sexing with Percoll Density Gradient Centrifugation (PDGC) method can separate X and Y spermatozoa based on their size (Rustidja, 1999). The study analyzed the results of X and Y spermatozoa sexing on Pangasius using the PDGC method and specified the ability of pH in influencing X and Y spermatozoa in the fertilization process. The results showed that, at stage 1, the spermatozoa head on the top layer (La) was larger than that on the bottom layer. The spermatozoa head length (HL) on the top layer was 2.59 ± 0.19 μm with a wide length (WL) of 1.62 ± 0.11 μm. Meanwhile, on the bottom layer, the HL was 2.27 ± 0.12 μm with a WL of 1.44 ± 0.07 μm. Therefore, the top layer was identified as X (female) and the bottom layer as Y (male). The average value of spermatozoa concentration after PDGC at the top layer was 7.11 ± 1.743 x 107 cells/mL and at the bottom layer was 3.57 ± 1.075 x 107 cells/mL. The average value of motility at the top layer was 61.67 ± 7.637% and at the bottom layer was 46.67 ± 5.773%, and the average value of viability at the top layer was 61.04 ± 7.27% and at the bottom layer was 55.17 ± 2.53%. This study revealed that the effect of pH on the fertilization rate (FR) with Y control spermatozoa was -177.8 + 68.377 x -4.234 x2, R² = 0.603. The finding confirmed that the best treatment for this control was pH 8.07 and FR 98.27%. The assessment on Y top layer spermatozoa resulted in of -100.27 + 40.836 x – 2.568 x2, R² 0.637. The finding showed that the best treatment for the top layer spermatozoa was pH 7.95 and FR 62.07%. The examination on Y bottom layer spermatozoa generated an equation of -32.195 + 22.746 x -1.4619 x2, R² = 0.523. It can be said that the best treatment in the bottom layer was pH 7.78 and FR 56.28%. However, the hatching rate (HR) to pH treatment on each treatment did not show significant differences (P >0.05). To sum up, the PDGC method on Pangasius spermatozoa could separate the spermatozoa based on the top and bottom layer size. The PDGC method could produce female seeds on the top layer by 85% and male seeds on the bottom layer by 68%.

In forensic anthropology, sex estimation is a fundamental step in assessing individualbiological profiles when analyzing human skeletons. Yet, current methods are notreliable enough to allow an accurate sex identification of highly fragmented, burnt,or subadult remains. This paper aims to investigate sexual dimorphism of the bonylabyrinth on both identified subadult and adult individuals. The bony labyrinth is ofparticular interest for sex estimation since it is alleged to complete size and maturationpre-pubertally and is located inside the petrous part of the temporal bone whichprotects it from taphonomic processes. The study was performed on 93 CT scans ofidentified individuals from two Belgian osteological collections (19–20th century) andfrom current pediatric images (Erasme hospital, Brussels). Linear and angular measurementswere taken on 2D slices of right bony labyrinths. Intra- and interobservers errormeasurements were calculated. Statistical tests were used to unravel any morphologicalvariations between subadult and adult bony labyrinths and to highlight differencesbetween females and males, separately in subadults and adults. Linear discriminantfunctions were established by cross-validation and tested on an independent samplefrom Belgium. Some measurements were significantly different between subadultsand adults, and between females and males within both subadult and adult samples.Univariate functions achieved 72.7% in subadults and 68.4% in adults whereas multivariateequations increased accuracy respectively up to 84.9% and 78.4%. This studyentails promising results to design a sex estimation method suitable for fragmentedand/or subadult remains. Further metric approaches are needed to explore bony labyrinthsexual dimorphism., info:eu-repo/semantics/published

Gender determination in incubated eggs (in ovo) has the potential to substitute the highly discussed practice of culling male layer chicks. The aim of this study was to investigate the effect pictures have on peoples' preferences toward in ovo sexing at different stages of embryonic development and chick culling. For this purpose, an online survey was conducted with a representative sample of 482 respondents in Germany. A within-subject design with 2 choice experiments was used to investigate the influence pictures have on respondents' preferences and willingness to pay. The first-choice experiment contained plain text only; the second contained also pictures of a chick or the incubated eggs at the corresponding stages of development. Findings reveal that in ovo gender determination at each proposed day of incubation (d1, d4, and d9) was preferred to chick culling. In ovo screening on d1 and d4 was significantly preferred to d9. This preference for early gender determination increased significantly as a consequence to the provision of pictures. Results furthermore reveal that a high error rate of gender determination or the lack of a meaningful utilization of incubated eggs can decrease approval for in ovo gender determination to an extent, where no positive willingness to pay remains. Findings of this study are useful for stakeholders in poultry production when considering the implementation of in ovo gender determination as a morally admissible substitute to chick culling.

Books Reviewed: The Better Half: On the Genetic Superiority of Women by Sharon Moalem (Penguin, April 2020) The Gendered Brain: The New Neuroscience that Shatters the Myth of the Female Brain by Gina Rippon (The Bodley Head, February 2019) Gender Mosaic: Beyond the Myth of the Male and Female Brain by Daphna Joel and Luba Vikhanski (Hachette, September 2019) Sexing the Body: Gender Politics and the Construction of Sexuality by Anne Fausto-Sterling (Basic Books, June 2020)

One environmentally friendly method used to manage Bactrocera dorsalis (Hendel), a key agricultural pest of substantial economic importance, is the sterile insect technique (SIT). Nevertheless, several deficiencies related to this strategy impair the success of the SIT, including the inferior performance of released sterile males compared with wild males, which could be partly solved by the utilization of gut symbionts as probiotic dietary components. In this study, a culture-dependent method was used to isolate and characterize gut-associated bacterial species in adult B. dorsalis genetic sexing strain (GSS) males. In addition, three bacterial isolates from the Enterobacteriaceae family, namely, Enterobacter sp., Morganella morganii, and Moellerella wisconsensis, were used as supplements in larval and adult diets to assess their effects on the life-history traits of irradiated males. Consistent with many previous studies, Enterobacter spp. was shown to be beneficial, with some quality control indices, such as adult size, pupal weight, survival rate under stress and nutritionally rich conditions, and mating competitiveness, being significantly increased, while slight nonsignificant increases in emergence rate and flight ability were observed. Conversely, the M. morganii and M. wisconsensis strains both had negative effects on irradiated male fitness and mating competitiveness. Our results, in combination with those of earlier studies, can contribute to improving the effectiveness of SIT application by enhancing the different aspects of augmentative rearing and biological traits of pests under laboratory rearing conditions.

Background With the enactment of the Brazilian Law Arouca 11,794/2008 and Decree 6.899/2009, there has been an urgent need for changes in the processes related to animal experimentation in Brazil; in particular, there is a need for improvements in enhancements of the lab animal management. To improve the management capacity of the Lab animal facility of the Carlos Chagas Institute’s Laboratory Animals Science (LAS), BioterC software was developed and implemented in 2014 for tracking mouse laboratory colonies. Five years after the implementation of this software, we sought to analyze the information in the database originated from BioterC using the Exploratory Analysis Data methodology (EDA). This article aims to identify animal breeding patterns using a data mining tool (Data Science) with Python programming language. Results The results show that from September 2014 to June 2019, under the license IACUC number LW- 6/17, 15.106 animals were produced. The C57BL/6, BALB/c and Swiss strains were the most frequently produced strains. The distribution of births due to crosses between these strains showed a median of 6 to 10 animals, depending on the genetic homozygosis and heterozygosis of the animal. The median number of days of mating was 35 days. In the sexing period, the records reported a median of 19 days. A total of 393 requests for animals from internal and external laboratories were registered. It was noted that approximately half of the animals produced to meet the demand for orders were discarded. Of the 15,106 animals, 38% were requested for animal experimentation, 58% were discarded and 4% did not have an outcome recorded in the data. Conclusions This volume of data provides an initial view of the information retrieval capabilities contained in BioterC, allowing for unique breeding knowledge by installing laboratory animals.

Sexing monomorphic birds by DNA and morphometrics allows researchers to study behavioural differences between the sexes. We aimed to verify the utility of the carpal spur to sex the monomorphic Blacksmith Lapwing or to determine a more suitable method. The Blacksmith Lapwing is a widespread resident of central and southern Africa. We used biometrics and carpal spur features of 116 Blacksmith Lapwings we ringed at five South African sites in 2008–2018 that were DNA-sexed from blood samples. Males were on average larger than females in most morphometrics, but all these measurements largely overlapped. Spur length was the most sexually dimorphic trait, then body mass and tarsus length. Male lapwings use their wing spurs in combat, thus the spurs’ length and sharpness, and body size, are likely subject to intrasexual selection. The male’s longer tarsus may be related to his role in scraping out the ground nest. Our backward stepwise analysis produced the discriminant function D1 = 0.366 tarsus length - 27.541 to determine this species’ sex, which correctly sexed 66% of our sample. If we allowed a 5% misclassification rate then 70% of individuals remained unsexed. Thus we recommend DNA analysis for studies that must accurately sex all Blacksmith Lapwings in a sample. All of the DNA-sexed adult males had sharp ivory tipped spurs but 36% had a spur shorter than 13 mm, proposed previously as the cut-off value between sexes. We suggest that Blacksmith Lapwings with sharp ivory-tipped spurs of 14 mm or longer can be safely sexed as males to reduce the need for expensive DNA tests.

Sex ratio (ratio between the number of male and female individuals in a population) is an important factor contributing to the survival rates of the population in wild. Imbalanced sex ratio is often the cause of diminished survival rates of endangered species. The analysis of DNA collected through noninvasive methodologies effectively allows to monitor populations of rare species or species that are difficult to observe in the wild. Such method was used to determine the sex ratio in populations of the capercaillie (Tetrao urogallus) and black grouse (Lyrurus tetrix) in the Tatra National Park. We collected over 250 samples (mostly excrement) in the area of the Tatra National Park and identified species via microsatellite genotyping and determined the sex using fragments of the CHD gene. We found that the majority of the samples belonged to male specimens irrespective of the species suggesting a large deviation from an equal sex ratio in wild populations of capercaillie and black grouse in the Tatra National Park. Our results point out the fundamental contribution of genetic assessment of sex ration to monitor the viability of both species in the Tatra National Park.

Livestock industry is one of the important pillars for country economy. Sperm sexing has the potential of significantly improving the quality and quantity of production in the livestock industries. Over years several attempts, involving several different parameters of sperm cells, have been made to develop an efficient sperm sorting procedure. Till date, the only commercially available method for bovine sperm is flow cytometry. The disadvantages associated with this approach, however, points towards the need for developing a more non-invasive and cost efficient approach for separation of X and Y chromosome bearing spermatozoa. In this review paper, we attempt to highlight all the previously developed approaches, as well as other emerging immunological and proteomics based methods that might have the potential of being developed as a promising breakthrough approach for semen sexing. Furthermore, we discuss the advantages and applications of semen sexing in India and around the world.

Background A genetic sexing strain (GSS) is an essential component for pest control using the sterile insect technique (SIT). A GSS is developed using a combination of Y-autosome translocation and a selectable marker such as pupal color, resulting in heterozygous males and homozygous females that possess wild-type brown pupae (wp+) and mutant white pupae (wp) alleles, respectively. The genetic sexing Salaya1 strain developed for Bactrocera dorsalis was evaluated using a clean stream and scaled-up for subsequent production lines (e.g., initiation, injection, and release). Colony management under small- and large-scale conditions for long-term rearing may affect the sexing system, genetic background, and fitness performance of the strain. Routine monitoring was applied to study genetic stability, genetic variation, and male mating competitiveness. Results The percentage of recombinants was significantly different between males (wp) and females (wp+), ranging between 0.21–0.43% and 0.01–0.04%, respectively. Using 106 bands from six ISSR markers, the genetic backgrounds of two generations (F40 and F108) of the clean stream were found to be almost identical (0.960), and between those two generations and the wild population, the similarities were 0.840 and 0.800, respectively. In addition, the sterile males performed well in competitive mating with fertile females (Relative Sterility Index = 0.67 ± 0.13). The rates of fliers calculated from both clean and release streams were higher than 0.95. Regarding the fitness of the Salaya1 strain, the fertility and pupal recovery were similar in all production lines. The sex ratio (Male/Female) distortion was also recorded. Conclusions The Salaya1 strain reared at the mass-rearing facility retained its genetic stability, genetic variation, behavior (e.g., competitive mating and flight ability), and traits related to fitness for at least 10 consecutive generations. The filter rearing system is effective at minimising the selection pressure while maintaining the genetic background and fitness performances of the clean stream. These characteristics were stable throughout the production lines. In addition, the production efficiency is comparable among the different production lines and other similar types of GSSs.

Heterotis niloticus is an African species of Osteoglossiformes that presents biological peculiarities and zootechnical performances favorable for fish farming. However, the absence of a sexual dimorphism hinders the optimization of its reproduction in captivity and limits the understanding of its reproductive behavior. This study is aimed at developing a minimally invasive and reliable sexing method to detect vitellogenin (Vtg) in female plasma. A commercial sexing kit (Acobium, Montpellier, France) for Arapaima gigas—a phylogenetically sister species of H. niloticus—successfully identified only 20% of mature H. niloticus females. Enzyme-linked immunosorbent assays (ELISA) were carried out using three Vtg antibodies. The A. gigas Vtg1 antibody cross-reacted significantly with plasma dilutions of female H. niloticus ranging from 1:1000 to 1:10,000, but with relatively low intensity. The Vtg antibody from Osteoglossum bicirrhosum, another species of Osteoglossiformes, showed non-specific binding with the Vtg of H. niloticus female plasma. Finally, an antibody for H. niloticus Vtg developed in this study allowed us to differentiate the two sexes with plasma coating dilutions ranging from 1:1000 to 1:10,000. The results of the assay were validated by a proteomic approach showing that Vtg-targeted mass spectrometry analysis of H. niloticus blood protein extracts could be used to accurately determine the presence of Vtg in the plasma of mature females. The final validation of the ELISA technique using the H. niloticus Vtg antibody was confirmed by visual sexing of a significant number of blood-sampled fish gonads; 100% of the fish were correctly sexed by the ELISA method.

It's a difficult task for researchers to identify the gender of chicken eggs by non-destructive approach in the early of incubation, which not only could reduce the cost of incubation, but also could improve the welfare of chicks. Therefore, SPME/GC-MS has been applied to investigate its potential as a non-destructive tool for characterizing the differences of odor between male and female chicken eggs during early of incubation and even before hatch. The results showed that more volatiles were found in female White leghorn eggs during early of incubation and 6,10-dimethyl-5,9-undecadien-2-one, 6-methyl-5-hepten-2-one, nonanal, decanal, octanal, 2-nonen-1-ol, etc. were important for the distinction of male and female White leghorn eggs during E1-E9 of incubation. 2-ethyl-1-hexanol; octanal, nonanal, 2,2,4-trimethyl-3-carboxyisopropyl pentanoic acid isobutyl ester; 2-nonen-1-ol, cyclopropanecarboxamide, heptadecane were correlated with gender of unhatched White leghorn, Hy-line brown and Jing fen eggs, respectively. Moreover, sex-related volatiles have been strongly influenced by incubation process and egg breed, and to be related to steroid hormone biosynthesis. What's more, this study enables us to develop a new visual for ovo sexing of chicken eggs and advances our understanding of the biological significance behind volatiles emitted from chicken eggs.

The USA has fallen, its borders have dissipated, and Black and Brown peoples have risen to power. In this setting, this essay is a fictional, first-person story from the perspective of a cyborg hum...

Demographic data including characterizations of population sex and age composition are of fundamental importance for effective management, especially for numerically depressed Great Lakes populations of imperiled species such as lake sturgeon (Acipenser fulvescens). The goal of this paper was to extend a recently reported Acipenserid-derived PCR-based genotyping test to determine sex of lake sturgeon. We demonstrate that the recently reported AllWSex2 primers amplified the female sex-specific region in lake sturgeon of known sex, consistent with a ZZ/ZW mode of inheritance. Sanger sequencing of female lake sturgeon PCR products matched published sequences from female sterlet (Acipenser ruthenus), showing 100% query cover and 98% (63/64) identity. The ability to provide a rapid, cost-effective, and unambiguous determination of sex for lake sturgeon will allow managers to determine compositional estimates of sex ratios during any season, and for individuals at any age or size, which is of great utility for species characterized by delayed sexual maturity and lacking external sexual dimorphisms.

The purpose of this study was to characterize spermatozoa motility of sexed semen (X and Y) using albumin by diluents and antioxidant treatments. The material used in this study was fresh semen of Ongole Crossbreed (OC) bull with progressive motility > 70%. The sperm sexing methodology used albumin gradient of 5%, 10%, and 15%. The diluents used were CEP-2 and andromed with or without the addition of 1mM Glutathione as an antioxidant. The sexed semen was made into liquid semen and stored at 3-5°C. The motility was observed at day 0 (H0) and day 5 (H5). Motility was analyzed using SCA v.2.1. The parameters measured were total motility, progressive motility, VCL, VSL, VAP, LIN, STR, WOB, ALH, BCF and sperm hyperactive. The experimental applied was the factorial pattern 2 x 2. The first factor was the type of diluents (CEP-2 and Andromed), and the second factor was additional glutathione (with or without). Data was analyzed by general linear model of SPSS-IBM 24 program. Sexed semen with CEP-2 diluents showed better motility of spermatozoa than andromed in the upper layers (spermatozoa X) and lower layers (spermatozoa Y). The CEP-2 diluent had a big role to maintain motility, progressive motility and velocity spermatozoa during cold storage. The addition of Glutathione 1mM could support the motility of sexed spermatozoa at cold storage, especially for LIN, STR, and WOB values .

В статье приводятся результаты применения метода бесконтактного определения пола птиц по профильному абрису головы к редкому нелетающему пастушку из Новой Зеландии - такахе. Графический этап позволил выявить визуальные различия в профильных абрисах клюва самцов и самок. Достоверность найденных различий была подтверждена набором статистических приемов анализа, показавших почти 100-процентную эффективность предложенного метода. Here we present the results of applying the method of non-contact sexing of birds by the profile beak outlines to a rare flightless rallid from New Zealand - takahē. The graphic stage of the method made it possible to identify visual differences in the profile outlines of the beak of males and females. The reliability of the differences found was confirmed by a set of statistical analysis techniques that showed almost 100 percent effectiveness of the proposed method.

Apoptosis is a fundamental process for the elimination of damaged or unwanted cells, and is a key aspect of development. It is triggered by pro-apoptotic genes responding to the intrinsic pathway that senses cell stress or the extrinsic pathway that responds to signals from other cells. The disruption of these genes can therefore lead to developmental defects and disease. Pro-apoptotic genes have been studied in detail in the fruit fly Drosophila melanogaster, a widely-used developmental model. However, little is known about the corresponding genes in its relative D. suzukii, a pest of soft fruit crops that originates from Asia but is now an invasive species in many other regions. The characterization of D. suzukii pro-apoptotic genes could lead to the development of transgenic sexing strains for pest management. Here, we describe the isolation and characterization of the pro-apoptotic genes reaper (Dsrpr), head involution defective (Dshid) and grim (Dsgrim) from a laboratory strain of D. suzukii. We determined their expression profiles during development, revealing that all three genes are expressed throughout development but Dsrpr is expressed most strongly, especially at the pupal stage. Functional analysis was carried out by expressing single genes or pairs (linked by a 2A peptide) in S2 cell death assays, indicating that Dsgrim and Dshid are more potent pro-apoptotic genes than Dsrpr, and the lethality can be significantly enhanced by co-expression of two genes. Therefore, the binary or multiple expression of different pro-apoptotic genes can be considered to build an efficient transgenic sexing system in D. suzukii.

In the winters of 1995, 1996 and 1997, research was conducted in the Auckland Islands to evaluate the status of southern right whales inthe New Zealand sub-Antarctic. Whales were present in high concentrations each year (maximum count of 146 whales) in a small area onthe northeast side of the main island. Cow-calf pairs averaged 12% (range 9% to 14%) of the total population. Most cow-calf pairs weresighted resting at the surface (60%) or travelling (36%) and showed a strong preference for shallow ( < 20m depth) nearshore waters. Theratio of females to males, as determined by molecular sexing using biopsy samples, varied from 54% to 39% over the three years but didnot differ significantly from 1:1 in any year. Both males and females were found in varying group sizes, with the occurrence of social/sexualactivity predominant (85%) in groups of three or more whales. Most single whales were found resting (59%) and occasionally approachedthe research vessel (19%). A total of 217 individual whales have been photo-identified over the three years of this study, 24% of which wereresighted more than once in a season and approximately 15% of which were resighted in more than one year. The high density of whalesin Port Ross during winter months, the presence of cow-calf pairs, including newborns, and the frequency of social and sexual activityindicates that the Auckland Islands are a primary wintering habitat for southern right whales in New Zealand waters. However, the lowresighting rates within season and documented movement to nearby Campbell Island (290km) suggest that some whales are not residentin the Auckland Islands throughout the season. The rarity of right whales along the main islands of New Zealand and their apparent increasein numbers in the Auckland Islands suggests a major shift in habitat use from pre-exploitation times or the loss of a component of ahistorically sub-divided stock.

The Sterile Insect Technique (SIT) is a target-specific, biologically based method used to control invading or established populations of pestiferous tephritids. SIT entails the release of large numbers of mass-reared, sterilized males, with the goal of achieving sterile male x wild female matings, which yield infertile eggs and suppress the growth rate of the pest population. The development of male-only strains generates the potential for more effective control of invading populations than standard bisexual releases. A genetic sexing strain based on sex-linked pupal color exists for Bactrocera dorsalis (Hendel), a major agricultural pest worldwide, but how this strain might perform in an SIT program has not been thoroughly investigated. As documented for several tephritid species, artificial selection imposed via mass-rearing may have adverse effects on various biological parameters of the released flies, including dispersion, life span, and mating competitiveness. The objective of the present study was to directly compare lure responsiveness using methyl eugenol, dispersal, and survival between males from wild and genetic sexing strains (GSS) of B. dorsalis. We conclude that males of the two strains differed significantly in dispersal ability but not in lure attraction or survival ability. The prospective effectiveness of the GSS in SIT programs against B. dorsalis is evaluated based on these findings.

The genetic sexing strain (GSS) of the Mediterranean fruit fly (Ceratitis capitata (Wiedemann)) Vienna 8D53− is based on a male-linked translocation system and uses two selectable markers for male-only production, the white pupae (wp) and the temperature sensitivity lethal (tsl) genes. In this GSS, males emerge from brown pupae and are resistant to high temperatures while females emerge from white pupae, are sensitive to high temperatures. However, double homozygous females (wp tsl/wp tsl) exhibit a slower development rate compared to heterozygous males (wp+tsl+/wp tsl) during the larval stage, which was attributed to the pleiotropic effects of the tsl gene. We present the first evidence that this slower development is due to a different gene, here namely slow development (sd), which is closely linked to the tsl gene. Taking advantage of recombination phenomena between the two loci, we report the isolation of a novel temperature sensitivity lethal strain using the wp mutation as a morphological marker, which showed faster development (wp tsl FD) during the larval stage and increased in its temperature sensitivity compared with the normal tsl strain. Moreover, the introgression of this novel wp tsl FD combined trait into the Vienna 8D53− GSS, resulted in a novel Vienna 8D53− FD GSS, where females showed differences in the thermal sensibility, larval development speed, and productivity profiles. The modification of these traits and their impact on the mass rearing of the GSS for sterile insect technique applications are discussed.

After suffering a massive decline (~ 99%) in numbers caused by feeding on livestock carcasses containing the nephrotoxic drug diclofenac, critically endangered Gyps vultures now persist in low numbers in the Indian subcontinent. Most are concentrated near National Parks, likely due to availability of wild carcasses free from toxic veterinary drugs. Quantification of vulture diets is critical to test this hypothesis. We describe a validated “field-to-benchtop-to-desktop” metabarcoding workflow for assessing species- and sex-specific diet of these obligate scavengers from non-invasively collected faecal samples. Seven metabarcodes were designed to simultaneouslsy identify the vulture species, sex and diet. Amplicons generated from multiplex PCRs were indexed and sequenced on an Illumina Miseq platform. We included controls and three replicates per sample to establish a series of non-arbitrary thresholds to filter the sequence data and eliminate cross-contamination, PCR/sequencing errors and false positives. This strategy enabled identification of species and sex for all samples and diet-specific sequences with high taxonomic resolution for 97% of samples. Just four (two for diet, one each for vulture species and sexing) metabarcodes were sufficient to meet the objectives. This preliminary analysis suggests that domestic livestock is the most frequently consumed diet item across samples from inside and outside protected habitats. Our method provides a rapid and reliable tool for describing large-scale variation in consumption of domestic versus wild species in the diet of these scavengers, paving the way for a better understanding of the role protected areas play in persistence and recovery of the remaining Gyps vultures in the wild.

Division of labor is fundamental to the ecological success of social insects. In termites, both sexes engage in social tasks, and the sexual division of labor is common in many taxa. Each caste consisting of both sexes is supplied from the population of newly hatched larvae. To understand their social system, it is necessary to investigate the influence of larval sex composition on their developmental fate. However, no method currently exists for sexing young larvae non-invasively, which is essential for experimental manipulation of the sex composition in a society. Here, we report on sex-specific characteristics of the first and second instar larvae of the subterranean termite Reticulitermes speratus. Male larvae possess bristles near the center of the posterior margin of the eighth abdominal sternite that are absent in females. The bristles are detectable under a stereomicroscope without damaging the young instar larvae. The validity of morphological sexing was confirmed by a known sex-specific genetic marker. The sex-specific bristles were also useful to identify the sex of first and second instar larvae of the damp-wood termite Zootermopsis nevadensis, suggesting that morphological sexing is possible for a wide range of termite species. The morphological sexing presented here has broad applicability in studies addressing sex differences in development, caste differentiation, and behavior. These approaches will contribute to understanding why both sexes co-exist in a termite society and allocate tasks, and which tasks are completely compensable by the other sex, thereby deepening our understanding of social systems where both sexes engage in tasks.

With the increasing availability of sexed semen, farms have the opportunity to select genetically superior dams to produce their replacement animals and to produce crossbred calves for beef production of higher economic value than the remainder of the herd. However, higher costs and reduced fertility of sexed semen complicate the decision of when and to what extent sexed semen should be applied in a herd. The objective of this study was to explore the economically optimal utilization of sexed semen and crossbreeding among North Rhine-Westphalian dairy farms in a holistic single-farm model. For the analysis, we derived a representative sample of farms from Latin Hypercube sampling based on the observed distribution of farm characteristics from official North Rhine-Westphalian Farm Structure Survey data. Market- and technology-related input parameters such as output prices and sexed semen accuracy and fertility were included in the sampling procedure. Modeling results of the systematic sensitivity analysis were evaluated in a statistical meta-model. We found that the profit-maximizing utilization of sexed semen and crossbreeding was highly heterogeneous among the farms. Farms with lower stocking densities,2 livestock units (LU)/ha, were generally found to produce excess heifers for sale, whereas farms with stocking densities2 LU/ha were producing crossbred calves and using sexed semen only to produce replacement animals. On average, female-sexed dairy semen was used on 25.3% of all inseminations. Beef semen (both sexed and conventional) for producing crossbred calves was used in an average of 21.5% of the inseminations. The combination of sexed semen and crossbreeding increased profits from €0 to €568 per cow per year, with an average of €79.42 per cow per year. Farms characterized by low stocking densities (2 LU/ha) and above-average replacement rates (40%) were found to have higher profit increases as a result of selling more heifers from the use of sexed semen. Overall, sexed semen and crossbreeding adoption were most sensitive to stocking density and average cow longevity, as well as to additional costs for sexed semen and sexed semen accuracy. Our results show the potential of modern breeding technologies to improve dairy farm profits and the need to judge their profitability in the light of farm-specific production settings.

Preimplantation genetic testing (PGT) routinely requires biopsy which is an invasive approach. The aim of this study was to examine a noninvasive approach for sexing of preimplantation embryos using polymerase chain reaction (PCR)/reverse transcriptase-PCR (RT-PCR) based on the presence of SRY DNA/RNA in the spent culture medium. Two groups were evaluated: in group 1, 40 embryos of routine PGT volunteers were cultured individually after biopsy and in group 2, 31 embryos were cultured individually until Day-5 post-fertilization. Each group was further divided into three subgroups: RNA extraction (RE), nucleic acid (NA) and DNase treated (DT). In the NA and DT subgroups, cDNA synthesis was performed directly on culture medium with or without DNase treatment in DT and NA subgroups, respectively. The results of sexing based on the PCR/RT-PCR in the culture medium, were compared with the results of sexing by fluorescence in situ hybridization (FISH) technique. In group 1, all samples were correctly diagnosed. In group 2, five female samples were misdiagnosed. Test's sensitivity, specificity and accuracy were 100 %, 94.44 % and 96.88 %, in RE, 100 %, 81.82 % and 93.55 % in DT and 100 %, 71.43 % and 85.71 % in NA, respectively. Preimplantation sexing without embryo biopsy, in the spent embryo culture media using RNA amplification based methods including RE and DT seem to be more reliable while nucleic acid based method, NA, led to the highest misdiagnoses probably due to DNA contamination. Since all male samples were correctly diagnosed in all subgroups of this preliminary study, preimplantation noninvasive sexing on culture medium seems feasible, however all sources of nucleic acid contamination must be carefully avoided.

The difficulty in differentiating the sex of monomorphic bird species has made molecular sexing an important tool in addressing this problem. This method uses noninvasively collected materials such as feathers and may be advantageous for sexing endangered as well as commercialized bird species. In this study, seven primer sets for sexing birds were screened in Aerodramus fuciphagus using a total of 13 feather samples that were randomly selected from the state of Perak, Malaysia. From the screening analysis, only one primer set (P8/WZ/W) successfully differentiated the sex of A. fuciphagus. PCR amplification produced a single 255‐bp DNA fragment for males which was derived from CHD‐Z (CHD gene region in the sex chromosome Z), while for the females it produced two fragments (144 and 255 bp). The 144‐bp fragment was from CHD‐W (CHD gene region in the sex chromosome W). Results from sequencing showed no variations in the base sequences of the CHD‐W and CHD‐Z amplified fragments within the same sexes, except for one male sample (A23) where at position 166, a base substitution occurred (G → A). Phylogenetic analysis of CHD‐W showed that four (Apodiformes; Gruiformes; Passeriformes; and Pelecaniformes) out of the five orders investigated had formed four clear clusters within their orders, including the studied order: Apodiformes. Whereas in CHD‐Z, four (Accipitriformes; Columbiformes; Galliformes; and Passeriformes) out of five orders investigated formed four clear clusters within their orders, excluding the studied order. In addition, A. fuciphagus and Apus apus (both Apodiformes) showed less divergence in CHD‐W than CHD‐Z (0% c.f. 9%). The result suggests that in A. fuciphagus, CHD gene evolution occurred at a higher rate in males (CHD‐Z) compared to females (CHD‐W). This finding may be useful for further studies on sex ratio and breeding management of A. fuciphagus., In this study, a molecular sexing method was used to identify the sex of monomorphic and economically important bird species; Aerodramus fuciphagus (Thunberg, 1812). Interestingly, we found one male sample (A23) where at position 166, had a base substitution (G → A), hence, an evolution study in CHD‐W and CHD‐Z genes by comparing A. fuciphagus with 14 other bird species from the same Apodiformes order (Apus apus and Calypte costae) and Aves class was performed. This study is important for further studies on sex ratio and breeding management of A. fuciphagus as well as CHD gene evolution.

The Mediterranean fruit fly Ceratitis capitata is a globally invasive pest, often controlled with the sterile insect technique (SIT). For the SIT, mass-rearing of the target insect followed by irradiation are imperatives. Sterile males are often less able to inhibit female remating and transfer less number of sperm, and even irradiation could affect male reproductive organs, with consequences for their ability to inhibit female remating. On the other hand, male age could affect their ability to modulate female response after mating. Here, we evaluated the quality of the genetic sexing strain Vienna-8-tsl mass-reared in Bioplanta San Juan, Argentina, under laboratory conditions, with regard to: (i) the ability of sterile males irradiated at 100 or 140 Gy to inhibit female remating, in the same day and at 24 h of first copulation; (ii) the ability of 3, 4 or 5 day-old sterile males to inhibit female remating at 24 h of first copulation, and (iii) the effect of a reduction in irradiation doses on the number of sperm stored by females and reproductive organ size in virgin males. Sterile males were better able than wild males to inhibit female remating in the same day of first copulation and as able as wild males 1 day after first copulation. Male age did not affect their ability to inhibit female receptivity. Number of sperm stored by females, testes size and ectodermal accessory glands size were not affected by male identity, while sterile 100 Gy males had larger mesodermal accessory glands than control lab males. A reduction in irradiation dose does not impact any variable measured, except for percentage of sperm-depleted females: females mated with sterile 100 Gy males had lower probabilities to store sperm. The results showed here are very encouraging for tsl Vienna 8 strain reared in Argentina and are discussed in comparison with previous studies in C. capitata female remating with dissimilar results.

This article summarises the surgical and non-surgical options for desexing dogs, using standardised terminology by defining the meaning of commonly used terms. Desexing has been historically recommended to help with population control, disease prevention and behaviour modification. Surgical methods of gonadectomy are the most common method for desexing in both owned and unowned dogs, with orchiectomy being the most common method of desexing male dogs. Surgical methods of desexing in females may refer to ovariohysterectomy, ovariectomy, salpingectomy or hysterectomy with or without an ovarian autograft (referred to as a so-called ‘ovary-sparing spay’). Non-surgical desexing methods include three broad categories: hormonal, immunological, and chemical or physical methods. The use of a gonadotropin-releasing hormone agonist as a method of inducing reversible infertility in dogs is discussed in detail, including a review of the physiological effects of surgical gonadectomy compared to a gonadotropin-releasing hormone agonist and potential health effects of each. There is a growing body of literature on the disease risks associated with whether a dog is intact or desexed, and by which method it has been desexed. This will be discussed in the next article.

This article presents an elaboration of the protocol for the method of sexing wild birds based on the polymorphism of the CHD gene using P2/P8 primer for Common Pheasant – Phasianus colchicus (Linnaeus, 1758; Galliformes, Phasianidae); Silver Lofur or Silver Pheasant – Lophura nycthemera (Linnaeus, 1758; Galliformes, Phasianidae), Budgerigar – Melopsittacus undulatus (Shaw, 1805; Psittaciformes, Psittacidae), Herring Gull – Larus argentatus (Pontoppidan, 1763; Charadriiformes, Laridae), and White Stork – Ciconia ciconia (Linnaeus, 1758; Ciconiiformes, Ciconiidae). Blood samples were taken from Common Pheasant, Silver Pheasant and White Stork using the “drop of blood on paper” method. For the Budgerigar and the Herring Gull, DNA was isolated from the feather follicle. To isolate DNA, a commercial NeoPrep 100 DNA reagent kit (Neogen, Ukraine) was used. Primers P2/P8 were used for PCR; PCR was performed using GenPac PCR Core reagents (Neogen, Ukraine). We selected the optimal amount of Tag polymerase, the amount of DNA and primers and, according to the amount of reagents, set acceptable amplification modes and electrophoresis agarose gel percentage. Prior to PCR, additional DNA gel electrophoresis purification is proposed, which increases the percentage of positive sex determination results. It was found that the ideal mixture for the 5 bird species was an amplification mixture (total volume 20 µL, containing 1 U Tag polymerase, 100 ng DNA and 0.6 µM of each primer). The amplified CHD-Z fragment of Common and Silver pheasants is of ~340 n. p., CHD-W ~360 n. p. Herring Gull and Budgerigar have ~350 n. p. of CHD-Z length, and ~400 n. p. of CHD-W length, White Stork has its CHD-Z of ~ 370 n. p. long. It is advisable to investigate the genome of the experimental bird species using horizontal electrophoresis in agar’s gel with the concentration of 5%, which makes it possible to clearly visualize the female genotype. The universal protocol of the method of sex determination based on polymorphism of the CHD gene for the 5 studied bird species is described. These results of the study led to the conclusion that for the simultaneous sexing of several species of birds, it is advisable to develop a unified protocol for determining the status of the CHD gene, with the aim of clarifying the gender, as well as new approaches in ornithology and ecology aimed at determining interspecific differences associated with gene polymorphism. Identification of differences in fragment sizes may be useful for identifying the species in cases when birds form mixed pairings for taxonomic and phylogenetic comparisons.